IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. J. Mol. Biol. (0022-2836) 2006 Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu Accepted 2007-JAN-23 by JYOTI. J. Mol. Biol. (0022-2836) 2006 A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. J. Mol. Biol. (0022-2836) 2006 Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu in vitro In vitro light scattering light scattering predetermined predetermined participant predetermined Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis Dimerization identified by mulit-light scattering using a 18-angle light scattering detector (DAWN EOS) and refractive index detector (Optilab DSP) (Wyatt Technology) kuriyan@berkeley.edu Accepted 2007-JAN-23 by JYOTI. J. Mol. Biol. (0022-2836) 2006 A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. J. Mol. Biol. (0022-2836) 2006 Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu Accepted 2007-JAN-23 by JYOTI. J. Mol. Biol. (0022-2836) 2006 A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases. J. Mol. Biol. (0022-2836) 2006 Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Pirruccello M., Sondermann H., Pelton JG., Pellicena P., Hoelz A., Chernoff J., Wemmer DE., Kuriyan J. Apoptosis - Interactions involving proteins with a function related to apoptosis kuriyan@berkeley.edu Accepted 2007-JAN-23 by JYOTI. J. Mol. Biol. (0022-2836) 2006 magnesium(2+) Magnesium(2+) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pak2_human Serine/threonine-protein kinase PAK 2 p21-activated kinase 2 Gamma-PAK PAK65 S6/H4 kinase p58 PAK2 protein protein human Homo sapiens Human MSDNGELEDKPPAPPVRMSSTIFSTGGKDPLSANHSLKPLPSVPEEKKPRHKIISIFSGTEKGSKKKEKERPEISPPSDFEHTIHVGFDAVTGEFTGMPEQWARLLQTSNITKLEQKKNPQAVLDVLKFYDSNTVKQKYLSFTPPEKDGFPSGTPALNAKGTEAPAVVTEEEDDDEETAPPVIAPRPDHTKSIYTRSVIDPVPAPVGDSHVDGAAKSLDKQKKKTKMTDEEIMEKLRTIVSIGDPKKKYTRYEKIGQGASGTVFTATDVALGQEVAIKQINLQKQPKKELIINEILVMKELKNPNIVNFLDSYLVGDELFVVMEYLAGGSLTDVVTETCMDEAQIAAVCRECLQALEFLHANQVIHRDIKSDNVLLGMEGSVKLTDFGFCAQITPEQSKRSTMVGTPYWMAPEVVTRKAYGPKVDIWSLGIMAIEMVEGEPPYLNENPLRALYLIATNGTPELQNPEKLSPIFRDFLNRCLEMDVEKRGSAKELLQHPFLKLAKPLSSLTPLIMAAKEAMKSNR DIP protein Q13177 original sequence version: 166 DIP protein Q13177 original sequence version: 114 00A7CD15F93D4180 limk1_human LIM domain kinase 1 LIMK1 LIMK protein protein human Homo sapiens Human MRLTLLCCTWREERMGEEGSELPVCASCGQRIYDGQYLQALNADWHADCFRCCDCSASLSHQYYEKDGQLFCKKDYWARYGESCHGCSEQITKGLVMVAGELKYHPECFICLTCGTFIGDGDTYTLVEHSKLYCGHCYYQTVVTPVIEQILPDSPGSHLPHTVTLVSIPASSHGKRGLSVSIDPPHGPPGCGTEHSHTVRVQGVDPGCMSPDVKNSIHVGDRILEINGTPIRNVPLDEIDLLIQETSRLLQLTLEHDPHDTLGHGLGPETSPLSSPAYTPSGEAGSSARQKPVLRSCSIDRSPGAGSLGSPASQRKDLGRSESLRVVCRPHRIFRPSDLIHGEVLGKGCFGQAIKVTHRETGEVMVMKELIRFDEETQRTFLKEVKVMRCLEHPNVLKFIGVLYKDKRLNFITEYIKGGTLRGIIKSMDSQYPWSQRVSFAKDIASGMAYLHSMNIIHRDLNSHNCLVRENKNVVVADFGLARLMVDEKTQPEGLRSLKKPDRKKRYTVVGNPYWMAPEMINGRSYDEKVDVFSFGIVLCEIIGRVNADPDYLPRTMDFGLNVRGFLDRYCPPNCPPSFFPITVRCCDLDPEKRPSFVKLEHWLETLRMHLAGHLPLGPQLEQLDRGFWETYRRGESGLPAHPEVPD DIP protein P53667 original sequence version: 113 9838BEFBE7006447 raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF DIP protein P04049 original sequence version: 117 DIP protein P04049 original sequence version: 106 DIP protein P04049 original sequence version: 114 DIP protein P04049 original sequence version: 141 DIP protein P04049 original sequence version: 142 DIP protein P04049 original sequence version: 147 DIP protein P04049-1 original sequence version: 136 DIP protein P04049 original sequence version: 136 DIP protein P04049 original sequence version: 153 DIP protein P04049 original sequence version: 150 DIP protein P04049 original sequence version: 164 DIP protein P04049 original sequence version: 172 DIP protein P04049-1 original sequence version: 172 DIP protein P04049 original sequence version: 184 DIP protein P04049 original sequence version: 188 DIP protein P04049 original sequence version: 195 EF821B5349711BC3 atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pak2-limk1-1 3 6 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range kinase region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q13177:p.Leu449Gln mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain L Q Wed May 02 17:00:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu449gln' Tue Jun 30 19:45:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu449Gln' ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 13 17 direct interaction direct interaction We noticed a dramatic change in the spectral properties of the kinase domain upon the addition of a large molar excess of LIMKtide (Figure 2(b)) with smaller line widths, an increase in intensity of weak peaks, and the appearance of new peaks. Fig 2 and Fig 6C pak2-raf1-1 3 6 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range kinase region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 8 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain in vitro In vitro 21 24 direct interaction direct interaction The addition of RAFtide but not PYE causes an improvement in line width and the appearance of new peaks. 2C pak2-pak2-1 2 6 unspecified role unspecified role neutral component neutral component region (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q13177:p.Leu449Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L Q Wed May 02 20:11:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu449gln' Tue Jun 30 19:09:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu449Gln' ecobb Escherichia coli (strain B / BL21) physical association physical association The phosphorylated PAK2Inact kinase domain dimerizes in solution. 3A, Table 1 pak2-pak2-2 4 6 unspecified role unspecified role neutral component neutral component kinase region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q13177:p.Leu449Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L Q Wed May 02 19:45:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu449gln' Tue Jun 30 15:40:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu449Gln' region (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) physical association physical association The phosphorylated PAK2Inact kinase domain dimerizes in solution. 3B, Table 1 Kd of L449Q mutant = 3.24E-4M pak2-atp-1 3 5 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range kinase region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) physical association physical association pak2-pak2-3 1 6 enzyme target enzyme target substrate substrate unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range kinase region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q13177:p.Leu449Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L Q Wed May 02 17:04:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu449gln' Tue Jun 30 18:12:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu449Gln' ecobb Escherichia coli (strain B / BL21) 6 enzyme enzyme unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation Enzyme target is catalytically inactive To test this, we performed kinase assays to measure the rate of incorporation of 32P-labeled phosphate onto the inactive kinase domain (PAK2Inact) by full length wild-type PAK2. 8B & 8C pak2-limk1-2 1 7 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 6 enzyme enzyme unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation The determination of KM values for (a) LIMKtide and (b) RAFtide. Initial velocities were measured with 40nM full length PAK2 using a coupled kinase assay Supp Fig 1 Km=2x10E-4M pak2-raf1-2 1 6 enzyme enzyme unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 8 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain in vitro In vitro phosphorylation phosphorylation reaction phosphorylation The determination of KM values for (a) LIMKtide and (b) RAFtide. Initial velocities were measured with 40nM full length PAK2 using a coupled kinase assay Supp. Fig. 1 Km=8.5x10E-5M